COYOTES AS A RESERVOIR FOR THE EMERGING ZOONOTIC PARASITE,

ONCHOCERCA LUPI, IN THE SOUTHWESTERN UNITED STATES

Chandler Roe, MS

Research Coordinator, PMI

Graduate Student, NAU

Chandler.Roe@nau.edu

April 2018

ONCHOCERCIASIS

• Filarial nematode

• Eye and skin disease

• O. volvulus: African River blindness

• Transmission: black fly bit, Simulium tribulatum

• 114 million treated in 2015

• Adult worms live ~15 years in host

• Female worms: 13-19 inches

• Male worms: 7-16 inches

• Wolbachia endosymbiont

WHO/TDR/stammers WHO/TDR/OCP

ONCHOCERCA VOLVULUS DISTRIBUTION 2015

1

Based on a figure from CDC

1

2

Based on a figure from CDC

1

2

3

Based on a figure from CDC

1

2

3

4

Based on a figure from CDC

1

2

3

4

5

Based on a figure from CDC

1

2

3

4

5

6

Based on a figure from CDC

ILLUSTRATIVE NUMBERS ON ONCHOCERCIASIS

Variable Value

Females per nodule 2 – 50 worms a

Males per nodule (constantly exchanging) 1 – 10 worms b

Microfilariae produced per day per adult female 1600

Total body daily microfilariae turnover (steady state) 10,000 – 300,000

Total body loads As high as 150 million microfilariae

a females remain incarcerated in nodulesb Males move in and out of nodules throughout lifespan

Udal 2007, CID

Heartworm

Filarial nematodes

Lefoulon et al 2017

wolf

WYATT EARPWyatt EarpRescue from Northern Arizona ReservationShotgun woundsVenereal diseasesEye infection in both eyesBroken tail

ONCHOCERCA LUPI

• Found: Northern AZ reservation

• Adopted: January 17, 2009

• Traveled:

• Yuma: December 2009

• Lee’s Ferry: March 2010

• New eye infection symptoms:

August 2010

Found single worm, sent off for morphological identification

Onchcocerca lupi

TREATMENT

Surgical excision of nodules

Melarsomine: An organic arsenical compound injected into the dog’s lumbar muscles.

Doxycycline: Antibiotic targets Wolbachia endosymbiont, sterilizes adult worms

Ivermectin: Anti-parasite medication

Prednisone: Anti-inflammatory steroid

Lots of eye drops..

May 2016

REPORTED ZOONOTIC O. LUPI IN HUMANS IN THE UNITED STATES

Age Case 122 months

Case 210 ya

Case 350 ya

Case 413 ya

Case 55 ya

Case 610 ya

Case 711 ya

Residence Arizona New Mexico Arizona Arizona New Mexico Texas Arizona

Location of nodule

Cervical spine Scalp Forearm Cervical spine Cervical spine Superior rectus muscle

Ocular

Method confirming diagnosis

Histology Histology, PCR

Histology, PCR

Histology Histology Histology Histology, PCR

Histologic findings

Gravid adult Nongravidadult

Nongravidadult

Nongravidadult

Gravid adult Multiple nongravid

adults

Unknown

Management Biopsy,ivermectin

Noduleexcision

Nodule excision,

ivermectin, doxycycline

Partial excision,

ivermectin, doxycycline

Nodule excision,

ivermectin, doxycycline

Nodule excision,

ivermectin, doxycycline

Unknown

ARIZONA CASE #1: 22 MONTH OLD FROM NORTHERN AZ

Eberhard et al, 2013

ARIZONA CASE #2

• 50 year old female, Southern Arizona

• Nickel-sized subcutaneous nodule on right forearm (November 2013)

• Rubbery, nontender, non-erythematous, and non-pruritic

• Travel to Monument Valley in Moab, Utah (2008)

• Subcutaneous granulomatous cyst removed (May 2014)

• Cyst contained O. lupi

Cross-section of adult female worm.Cantey, P. et. al. 2016. Clinical Infectious Diseases.

ARIZONA CASE #3

• 13 year old male, NE Arizona

• Week + worsening left-sided neck pain, sore throat, dysphagia, and headache

• Treatment with anti-inflammatories and antibiotics

• Worsening pain/headache 2 days later, with meningismus

• CSF evaluation revealed bacterial meningitis

• Treatment Improvement

ARIZONA CASE #3 CONT’D

• Symptoms reoccurred 4 weeks later

• CT normal

• MRI showed intradural, extramedullary mass within upper cervical spinal canal

• O. lupi infection confirmed by CDC

• Ophthalmological examination normal

• Patient treated with 6-weeks of doxycycline

• Repeat MRI after showed decrease in size of mass

MRI cervical spine, Arizona case #3.Cantey, P. et. al. 2016. Clinical Infectious Diseases.

May 201222 monthsApache County

April 201413 yearsApache County

May 201450 yearsMaricopa County

2015-2017Coyote Project Implementation

July 201711 yearsCoconino County

Canines as suspected host and sentinels.

STUDY OBJECTIVES

• Characterize the presence and geographic distribution patterns of O. lupi in Arizona coyote populations

• Identify the role of coyotes as potential hosts of O. lupi and as a potential reservoir for domestic dog or human infections

• December 2015 - May 2017

• Sampling kits provided

• Skin tissue from base of ear

• Routine coyote removal operations

SAMPLE COLLECTION

603Coyotes

571Arizona

26New Mexico

6Nevada

SAMPLING DISTRIBUTION

0 100 200 300 400 500

Majority of coyotes sampled were adults.

Adult

Youth

0 100 200 300 400

Majority of coyotes sampled were male.

Male

Female

METHODS

• Tissue processing

• DNA extraction

• Real-Time PCR screening

• Amplicon sequencing

• Bioinformatics

U.S. strains are closely related.

Fairly new introduction of O. lupi in the U.S.

NEXT STEPS

• Surveillance

• Black flies• Humans

• Animals

• Expand study site

• Outreach

• Clinicians

• Veterinarians

• Diagnostic

• Collect blood from positive and negative canine samples

• Draft genome

• Sequence whole genome

• Better population genomics

COLLABORATORS

Hayley Yaglom

Dr. David Engelthaler

Dr. Jennifer Urbanz

Veronica Harrison

Ted Lyons

Dr. Jolene Bowers

Wyatt Earp

THANK YOU

Chandler Roe, MSChandler.Roe@nau.edu | 928-503-3280